Issue 55, 2020

Molecular crowding induces primer extension by RNA polymerase through base stacking beyond Watson–Crick rules

Abstract

The polymerisation of nucleic acids is essential for copying genetic information correctly to the next generations, whereas mispolymerisation could promote genetic diversity. It is possible that in the prebiotic era, polymerases might have used mispolymerisation to accelerate the diversification of genetic information. Even in the current era, polymerases of RNA viruses frequently cause mutations. In this study, primer extension under different molecular crowding conditions was measured using T7 RNA polymerase as a model for the reaction in the prebiotic world. Interestingly, molecular crowding using 20 wt% poly(ethylene glycol) 2000 preferentially promoted the primer extensions with ATP and GTP by T7 RNA polymerase, regardless of Watson–Crick base-pairing rules. This indicates that molecular crowding decreases the dielectric constants in solution, resulting in enhancement of stacking interactions between the primer and an incorporated nucleotide. These findings suggest that molecular crowding could accelerate genetic diversity in the prebiotic world and may promote transcription error of RNA viruses in the current era.

Graphical abstract: Molecular crowding induces primer extension by RNA polymerase through base stacking beyond Watson–Crick rules

Supplementary files

Article information

Article type
Paper
Submitted
27 jul 2020
Accepted
27 aug 2020
First published
07 sep 2020
This article is Open Access
Creative Commons BY-NC license

RSC Adv., 2020,10, 33052-33058

Molecular crowding induces primer extension by RNA polymerase through base stacking beyond Watson–Crick rules

S. Takahashi, H. Okura, P. Chilka, S. Ghosh and N. Sugimoto, RSC Adv., 2020, 10, 33052 DOI: 10.1039/D0RA06502A

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